@article{1822e110a2fa4621b59ac4a8835c1f0f,
title = "Yeast translation elongation factor eEF3 promotes late stages of tRNA translocation",
abstract = "In addition to the conserved translation elongation factors eEF1A and eEF2, fungi require a third essential elongation factor, eEF3. While eEF3 has been implicated in tRNA binding and release at the ribosomal A and E sites, its exact mechanism of action is unclear. Here, we show that eEF3 acts at the mRNA–tRNA translocation step by promoting the dissociation of the tRNA from the E site, but independent of aminoacyl-tRNA recruitment to the A site. Depletion of eEF3 in vivo leads to a general slowdown in translation elongation due to accumulation of ribosomes with an occupied A site. Cryo-EM analysis of native eEF3-ribosome complexes shows that eEF3 facilitates late steps of translocation by favoring non-rotated ribosomal states, as well as by opening the L1 stalk to release the E-site tRNA. Additionally, our analysis provides structural insights into novel translation elongation states, enabling presentation of a revised yeast translation elongation cycle.",
keywords = "ABC ATPase, E-site tRNA, L1 stalk, cryo-EM, eEF3",
author = "Namit Ranjan and Pochopien, {Agnieszka A.} and {Chih-Chien Wu}, Colin and Bertrand Beckert and Sandra Blanchet and Rachel Green and {V Rodnina}, Marina and Wilson, {Daniel N.}",
note = "Funding Information: We thank Prof. Ralf Ficner for providing the strains for eIF1 and eIF1A purification. We thank Theresia Steiger, Tessa H{\"u}bner, Olaf Geintzer, Susanne Rieder for expert technical assistance, Paul Huter for help with data processing, and Otto Berninghausen and Roland Beckmann (Gene Center, LMU, Munich) for data collection. This work has been supported by iNEXT (project number 2643 to D.N.W.), the Horizon 2020 program of the European Union (CEITEC MU), NIH (R37GM059425 to R.G.), HHMI (R.G and C.C.W.), and grants of the Deutsche Forschungsgemeinschaft (DFG) WI3285/8‐1 to D.N.W., RA 3194/1‐1 to N.R., and Leibniz Prize to M.V.R.. This article reflects only the author{\textquoteright}s view and the European Commission is not responsible for any use that may be made of the information it contains. CIISB research infrastructure project LM2015043 funded by MEYS CR is gratefully acknowledged for the financial support of the measurements at the CF Cryo‐electron Microscopy and Tomography CEITEC MU. Open Access funding enabled and organized by ProjektDEAL. Publisher Copyright: {\textcopyright} 2021 The Authors. Published under the terms of the CC BY 4.0 license",
year = "2021",
month = mar,
day = "15",
doi = "10.15252/embj.2020106449",
language = "English (US)",
volume = "40",
journal = "EMBO Journal",
issn = "0261-4189",
publisher = "Nature Publishing Group",
number = "6",
}